Biomedical subjects

W Zhang

Publications and source records attributed to W Zhang.

At least 127 records · Page 7Linked to original sources

PubMed2003

Protein-protein interactions involving inducible nitric oxide synthase.

AIM: Nitric oxide (NO) is a signaling and effector molecule that contributes to multiple physiological and pathophysiological processes in the kidney, vasculature, and other tissues. High output NO generation by inducible NO synthase (iNOS) participates in host defense against pathogens and contributes to tissue injury during inflammatory states. Because of its potent reactivity and diffusibility, NO generation by iNOS is subject to multiple levels of regulation, including transcriptional, translational, and post-translational controls, including protein-protein interactions. This review examines the experimental basis for these protein-protein interactions and their known and potential importance for kidney and vascular physiology. METHODS: Analysis of the biomedical literature in the area. RESULTS: iNOS interacts with the inhibitory molecules Kalirin and NOS-associated protein 1.10 kd (NAP110), which inhibit iNOS homodimerization, as well as activator proteins, the Rac-GTPases. Interactions with caveolin-1 control the intracellular locale and degradation of iNOS in tumor cells. In polarized epithelial cells, associations of iNOS with the scaffolding protein EBP50 position iNOS in the apical membrane near key ion transport proteins that also interact with EPB50. In addition, protein-protein interactions of proteins governing iNOS transcription function to specify activation or suppression of iNOS induction by cytokines. CONCLUSION: Interactions of iNOS with a diverse group of heterologous proteins provides a selective mechanism to control the activity, spatial distribution, and proximity of iNOS to intended targets, while potentially limiting autotoxicity to the iNOS-expressing cell.

Blood Proteins↗

PubMed2003

The role of the cytoskeleton in capacitative calcium entry in myenteric glia.

Capacitative calcium entry (CCE) is the process by which intracellular calcium is replenished from the external milieu upon depletion of intracellular stores. CCE is thought to participate in chemotaxis, proliferation and cell signalling. A physical interaction between intracellular stores and the plasma membrane is postulated to regulate CCE. We hypothesized that cytoskeletal disruption alters this interaction, inhibiting CCE in enteric glia. Cultured myenteric glia from neonatal guinea-pigs were treated with cytochalasin D (10 micro mol L-1), a microfilament disrupting agent, nocodazole (20 micro mol L-1), a microtubule disrupting agent, or vehicle (dimethyl sulphoxide). Intracellular calcium changes were measured using fura-2 microfluorimetry. To evaluate the rate of cation re-entry, barium was substituted for calcium because barium is not sequestered internally. Cytochalasin D-treated glia had diminished CCE responses (57 +/- 3 nmol L-1) compared with controls (97 +/- 7 nmol L-1) as did nocodazole-treated glia (30 +/- 2 nmol L-1) vs controls (77 +/- 6 nmol L-1). The proportion of cells demonstrating CCE abolition was greater in the cytochalasin (50 +/- 8%) and nocodazole-treated (89 +/- 2%) groups compared with controls (21 +/- 2%, 40 +/- 9%, respectively). Cytochalasin D and nocodazole treatment diminished the rate of cation re-entry based on diminished barium entry in treated vs control cells. From this study, we conclude that disruption of cytoskeletal elements diminishes calcium influx essential to calcium store repletion in myenteric glia.

Animals↗

PubMed2003

Glycoprotein IIb/IIIa inhibition attenuates platelet-activating factor-induced platelet activation by reducing protein kinase C activity.

Glycoprotein (GP)IIb/IIIa inhibition may abolish activated leukocyte-induced platelet activation, in which leukocyte-released platelet-activating factor (PAF) is a major mediator. The present study thus investigated if and how GPIIb/IIIa inhibitors interfere with PAF-induced platelet activation. Platelet and leukocyte activation were monitored by flow cytometry and immunoblotting. GPIIb/IIIa inhibitors (c7E3, non-peptide SR121566, and MAb RFGP56) attenuated PAF-induced, but not adenosine diphosphate (ADP)- or thrombin receptor activating peptide (TRAP)-induced platelet P-selectin expression in whole blood. GPIIb/IIIa blockade enhanced ADP- or TRAP-induced leukocyte CD11b expression, but not the response to PAF. GPIIb/IIIa blockade attenuated PAF-induced, but enhanced ADP- or TRAP-induced platelet-leukocyte aggregation. Under the present experimental conditions, thromboxane A2 receptor antagonism did not significantly influence PAF-induced platelet activation, and GPIIb/IIIa inhibition did not interfere with calcium mobilization/influx in platelets. Protein kinase C (PKC) blockade inhibited PAF-induced platelet P-selectin expression, and PAF-induced PKC activity was reduced by GPIIb/IIIa inhibition. PAF (=1 micro m) did not induce MEK 1/2 or ERK 1/2 phosphorylation, whilst thrombin induced marked responses, which were enhanced by GPIIb/IIIa blockade. Thus, GPIIb/IIIa inhibition attenuates PAF-induced platelet activation via inhibiting PKC activity. GPIIb/IIIa blockade enhances thrombin-induced platelet MEK 1/2 and ERK 1/2 activation, and augments ADP- and TRAP-induced leukocyte activation by enhancing platelet-leukocyte aggregation.

Adenosine Diphosphate↗

PubMed2003

Genetics of phytate phosphorus bioavailability: heritability and genetic correlations with growth and feed utilization traits in a randombred chicken population.

The current study was undertaken to estimate variance components for phytate P bioavailability (PBA) and the genetic correlations among PBA with growth and feed utilization (or intake) traits in an unselected random mating chicken population. Pedigreed data from 901 Athens-Canadian randombred chickens hatched from 26 sires, 71 dams, and 105 grandparents were used for estimation of genetic parameters. Birds were individually housed in metabolic cages at 4 wk of age and fed a 0.35% P diet. After 3 d of acclimatization, excreta produced for 3 consecutive d were collected and feed consumed (FC) was measured. Individual 4-wk BW and BW gain (BWG) during the 3-d excreta collection period were also measured. Feed conversion ratios (FCR) were calculated. Phytate P bioavailability was estimated from the disappearance of phytate during the passage of feed through the gastrointestinal tract. The restricted maximum likelihood method with the average information matrix algorithm was used for the estimation of variance components. The heritability estimate for PBA was about 0.10. Genetic correlations between PBA and BW, BWG, and FC were moderate and negative, indicating that improving PBA utilization would moderately affect growth. The genetic correlation between PBA and FCR was negligible and suggested that selection for PBA will not adversely affect FCR. The economic implications of genetically modifying poultry to improving phytate P utilization and the subsequent elimination or reduction of the amount of phytase used in poultry diets are yet to be determined.

Animal Nutritional Physiological Phenomena↗

PubMed2003

An involvement of trigeminal mesencephalic neurons in regulation of occlusal vertical dimension in the guinea pig.

Although the occlusal vertical dimension (OVD) is strictly controlled, the neuronal mechanism of its regulation is still unclear. We hypothesize that neurons in the trigeminal mesencephalic nucleus (MesV) play an important role in the regulation of the OVD, because the MesV receives the projection from jaw-closing muscle spindles and periodontal mechanoreceptors. We measured the temporal OVD change in the guinea pig to study the effects of MesV lesions on the OVD. OVD-raised animals without MesV lesions showed a rapid OVD decrease to the same level as that in naïve controls, followed by an OVD increase after the OVD-raising appliance was removed. In contrast, OVD-raised animals with MesV lesions showed only a slight decrease in the OVD for 15 days after removal of the appliance, and then the OVD increased. The time-course of OVD development in normal-bite animals with MesV lesions was similar to that of naïve controls. These results suggest that MesV neurons are involved in OVD regulation.

Animals↗

PubMed2003

Scenarios for autoimmunization of T and B cells in myasthenia gravis.

We have studied responses in thymoma patients to interferon-alpha and to the acetylcholine receptor (AChR) in early-onset myasthenia gravis (EOMG), seeking clues to autoimmunizing mechanisms. Our new evidence implicates a two-step process: (step 1) professional antigen-presenting cells and thymic epithelial cells prime AChR-specific T cells; then (step 2) thymic myoid cells subsequently provoke germinal center formation in EOMG. Our unifying hypothesis proposes that AChR epitopes expressed by neoplastic or hyperplastic thymic epithelial cells aberrantly prime helper T cells, whether generated locally or infiltrating from the circulation. These helper T cells then induce antibody responses against linear epitopes that cross-react with whole AChR and attack myoid cells in the EOMG thymus. The resulting antigen-antibody complexes and the recruitment of professional antigen-presenting cells increase the exposure of thymic cells to the infiltrates and provoke local germinal center formation and determinant spreading. Both these and the consequently enhanced heterogeneity and pathogenicity of the autoantibodies should be minimized by early thymectomy.

Age of Onset↗

PubMed2003

Modeling human breast cancer metastasis in mice: maspin as a paradigm.

Breast cancer is the most common cancer detected in women, accounting for nearly one out of every three cancers diagnosed in the United States. Most cancer patients do not die from the primary tumor but die due to metastasis. Therefore, the study of metastasis is of most importance both to the clinician and patient. In the past, animal models have been used in breast cancer research and mammary gland biology. Our group has also established several animal models to address the function of a novel tumor suppressor gene maspin in breast tumor progression. Maspin was initially isolated from normal mammary epithelial cells. Its expression was down regulated in breast tumors. To test the protective role of maspin overexpression in mammary tumor progression, we crossed maspin overexpression transgenic mice (WAP-maspin) with a strain of oncogenic WAP-SV40 T antigen mice. The bitransgenic mice had reduced tumor growth rate and metastasis. Maspin overexpression increased the rate of apoptosis of both preneoplastic and carcinomatous mammary epithelial cells. Maspin reduced tumor growth through a combination of reduced angiogenesis and increased apoptosis. In a separate animal experiment, maspin overexpressing mammary tumor cells (TM40D) were implanted into the fat pad of syngeneic mice. TM40D tumor cells were very invasive and metastatic. However, both primary tumor growth and metastasis were significantly blocked in TM40D cells that overexpress maspin as a consequence of plasmid or retrovirus infection. These evidences demonstrate that maspin function to inhibit primary tumor growth as well as invasion and metastasis. Elucidating the molecular mechanism of maspin action will shed light on our understanding of breast cancer invasion and metastasis.

Animals↗

PubMed2003

RNA amplification strategies for cDNA microarray experiments.

The biological materials available for cDNA microarray studies are often limiting. Thus, protocols have been developed to amplify RNAs isolated from limited amounts of tissues or cells. RNA amplification by in vitro transcription is the most widely used among the available amplification protocols. Two means of generating a dsDNA template for the RNA polymerase are a combination of reverse transcription with conventional second-strand cDNA synthesis and a combination of the switch mechanism at the 5' end of RNA templates (SMART) with reverse transcription, followed by PCR. To date, there has been no systematic comparison of the efficiency of the two amplification strategies. In this study, we performed and analyzed a set of six microarray experiments involving the use of a "regular" (unamplified) microarray experimental protocol and two different RNA amplification protocols. Based on their ability to identify differentially expressed genes and assuming that the results from the regular protocol are correct, our analyses demonstrated that both amplification protocols achieved reproducible and reliable results. From the same amount of starting material, our results also indicated that more amplified RNA can be obtained using conventional second-strand cDNA synthesis than from the combination of SMART and PCR. When the critical issue is the amount of starting RNA, we recommend the conventional second-strand cDNA synthesis as the preferred amplification method.

DNA, Complementary↗

PubMed2003

M1-M5 muscarinic receptor knockout mice as novel tools to study the physiological roles of the muscarinic cholinergic system.

A large body of evidence indicates that muscarinic acetylcholine receptors (mAChRs) play critical roles in regulating the activity of many important functions of the central and peripheral nervous systems. However, identification of the physiological and pathophysiological roles of the individual mAChR subtypes (M(1)-M(5)) has proven a difficult task, primarily due to the lack of ligands endowed with a high degree of receptor subtype selectivity and the fact that most tissues and organs express multiple mAChRs. To circumvent these difficulties, we used gene targeting technology to generate mutant mouse lines containing inactivating mutations of the M(1)-M(5) mAChR genes. The different mAChR mutant mice and the corresponding wild-type control animals were subjected to a battery of physiological, pharmacological, behavioral, biochemical, and neurochemical tests. The M(1)-M(5) mAChR mutant mice were viable and reproduced normally. However, each mutant line displayed specific functional deficits, suggesting that each mAChR subtype mediates distinct physiological functions. These results should offer new perspectives for the rational development of novel muscarinic drugs.

Animals↗

PubMed2002

MoeA, an enzyme in the molybdopterin synthesis pathway, is required for rifamycin SV production in Amycolatopsis mediterranei U32.

Rifamycin SV contains one amide nitrogen atom at its C(7)N moiety. Earlier labeling studies suggested that nitrogen might be incorporated from a pathway involved in a molybdenum-dependent nitrate reductase. However, no genetic evidence is available thus far. The structural gene moeA, which is involved in molybdopterin synthesis in various organisms, has been cloned from rifamycin SV-producing Amycolatopsis mediterranei strain U32. The amino acid sequence deduced from the moeA gene showed significant similarity to members of the MoeA protein family and contains all the structural features that are highly conserved in the putative functional domains of MoeA proteins. Southern hybridization showed that there is only one moeA gene in the A. mediterranei genome. To further investigate the possible physiological function of the moeA gene, a double crossover gene replacement was achieved by inserting an aparmycin resistance gene into moeA in the A. mediterranei U32 chromosome. Phenotype analysis showed that the moeA gene is required for A. mediterranei growth in a minimal medium with nitrate as sole nitrogen source, possibly because nitrate reductase activity is diminished due to disruption of the moeA gene. Compared to the wild type strain, moeA-disrupted mutants lost 95% of their rifamycin SV production capacity in complex fermentation media. The results demonstrate that the moeA gene is necessary for rifamycin SV production in A. mediterranei, and that the nitrogen assimilation pathway involved in nitrate reductase is the major pathway for the genesis of the amide nitrogen atom in the rifamycin SV molecule.

Actinomycetales↗

PubMed2002

Modeling for point-non-point source effluent trading: perspective of non-point sources regulation in China.

In the past decades, little abatement efforts have been implemented on China's non-point source water pollution, and studies aiming at non-point sources regulation were also rare. Watershed abatement trading between point and non-point sources may serve as a cost-effective way to deal with the problem. The inherent uncertainty of non-point emissions, however, could affect the feasibility and outcome of point-non-point effluent trading. The purpose of this paper is to model the watershed point-non-point abatement trading incorporating the uncertainty of non-point source emissions, and to examine its impacts on trading equilibrium and trading ratio. The uncertainties of non-point emissions were taken into consideration by setting an acceptable probability by which the watershed emission constraints were achieved. Using the watershed optimization model, the optimal abatement allocation and trading ratio were explicitly illustrated. It was found that they were affected significantly by the variances of non-point emissions, the reliability requirement assigned to the non-point abatement, and the marginal abatement costs of point and non-point sources. Since the variances of non-point emissions may increase or decrease at the abatement level, the impacts of these factors were discussed in different circumstances. Based on the illumination of the trading model, future directions and implications of point-non-point trading in China were discussed.

Journal Article↗

PubMed2002

Differential ontogeny of GABA(B)-receptor-mediated pre- and postsynaptic modulation of GABA and glycine transmission in respiratory rhythm-generating network in mouse.

Rhythm generation in mature respiratory networks is influenced strongly by synaptic inhibition. In early neonates, GABA(A)-receptor- and glycine-receptor-mediated inhibition is not present, thus the question arises as to whether GABA(B)-receptor-mediated inhibition plays an important role. Using brainstem slices of neonatal mice (postnatal day, P0-P15), we analysed the role of GABA(B)-mediated modulation of GABA and glycine synaptic transmission in the respiratory network. Blockade of GABA uptake by nipecotic acid (0.25-2 mM) reduced the respiratory frequency. This reduction was prevented by the selective GABA(B) receptor antagonist CGP55845A (CGP) alone at P0-P3, but by bicuculline as well as CGP at P7-P15. Blockade of GABA(B) receptors by CGP increased the respiratory frequency at P0-P3, whereas it caused a reduction of frequency in older animals. The effect of CGP on respiratory frequency was diminished in the presence of bicuculline and strychnine in older but not in younger animals. The relative contribution of GABA(B)-receptor-mediated pre- and postsynaptic modulation was examined by analysing the effect of GABA(B) receptors on spontaneous and miniature IPSCs. In younger animals (P0-P3), the GABA(B) receptor agonist baclofen had no detectable effect on IPSC frequency, but caused a significant decrease in the amplitude. In older animals (P7-P15), baclofen decreased both the frequency and amplitude of spontaneous and miniature IPSCs. These results demonstrate that GABA(B)-receptor-mediated postsynaptic modulation plays an important role in the respiratory network from P0 on. GABA(B)-receptor-mediated presynaptic modulation develops with a longer postnatal latency, and becomes predominant within the first postnatal week.

Aging↗

PubMed2002

Collapse of valence transition in Yb0.8Y0.2InCu4: pressure-induced weak ferromagnetism.

We investigated the magnetization of Yb0.8Y0.2InCu4 as a function of temperature down to 0.6 K, pressure up to 1.2 GPa, and magnetic field up to 9 T. The valence transition temperature of Yb0.8Y0.2InCu4 is lowered with applying pressure. At 0.8 GPa, collapse of the valence transition and ferromagnetic ordering occur almost simultaneously. The ferromagnetic phase at 1.2 GPa is characterized by a low Curie temperature of 1.7 K and an extremely small ordered moment of 0.05 micro(B) per Yb. Some effect of screening the ordered moment may play a key role in the ferromagnetism and the valence transition.

Journal Article↗

PubMed2002

Diffraction of a superfluid fermi gas by an atomic grating.

An atomic grating generated by a pulsed standing-wave laser field is proposed to manipulate the superfluid state in a quantum degenerate gas of fermionic atoms. We show that in the presence of atomic Cooper pairs, the density oscillations of the gas caused by the atomic grating exhibit a much longer coherence time than that in the normal Fermi gas. Our result indicates that the technique of a pulsed atomic grating is a potential candidate to detect the atomic superfluid state in a quantum degenerate Fermi gas.

Journal Article↗

PubMed2002

GCPII (NAALADase) inhibition prevents long-term diabetic neuropathy in type 1 diabetic BB/Wor rats.

AIMS/HYPOTHESIS: Hyperglutamatergic activity induced by ischemia is believed to underlie neuronal damage in a variety of neurological disorders, including neuropathic pain. Since ischemia is believed to be a prominent mechanism involved in diabetic polyneuropathy (DPN), we investigated the effect of the glutamate carboxypeptidase II (GCPII, EC #3.4-17.21; previously termed NAALADase), an enzyme responsible for the hydrolysis of the neuropeptide NAAG to NAA and glutamate, on the development of DPN in type 1 diabetic BB/Wor rats. METHODS: Diabetic animals were treated with 10 mg/kg/day i.p. of the selective GCPII inhibitor GPI-5232 from onset of diabetes for 6 months. Hyperalgesia to thermal stimulation and nerve conduction velocity (NCV) were measured monthly. The effect on structural DPN was assessed by scoring of single, teased myelinated fibers, myelinated fiber morphometry and ultrastructural examination of C-fibers at 6 months. RESULTS: GCPII inhibition showed significant but partial effects on hyperalgesia (p<0.001), nerve conduction slowing (p<0.01) axonal and nodal structural changes (p<0.001), small myelinated fiber atrophy, and degenerative changes of C-fibers. CONCLUSIONS: GCPII inhibition has beneficial effects on hyperalgesia, nerve function, and structural degenerative changes in DPN, which are likely mediated by inhibition of ischemia-induced glutamate release.

Animals↗

PubMed2002

Cellular lipid peroxidation end-products induce apoptosis in human lens epithelial cells.

Hydrogen peroxide (H(2)O(2)), an oxidant present in high concentrations in the aqueous humor of the elderly eyes, is known to impart toxicity to the lens---apoptosis being one of the toxic events. Since H(2)O(2) causes lipid peroxidation leading to the formation of reactive end-products, it is important to investigate whether the end-products of lipid peroxidation are involved in the oxidation-induced apoptosis in the lens. 4-Hydroxynonenal (HNE), a major cytotoxic end product of lipid peroxidation, has been shown to mediate oxidative stress-induced cell death in many cell types. It has been shown that HNE is cataractogenic in micromolar concentrations in vitro, however, the underlying mechanism is not yet clearly understood. In the present study we have demonstrated that H(2)O(2) and the lipid derived aldehydes, HNE and 4-hydroxyhexenal (HHE), can induce dose- and time-dependent loss of cell viability and a simultaneous increase in apoptosis involving activation of caspases such as caspase-1, -2, -3, and -8 in the cultured human lens epithelial cells. Interestingly, we observed that Z-VAD, a broad range inhibitor of caspases, conferred protection against H(2)O(2)- and HNE-induced apoptosis, suggesting the involvement of caspases in this apoptotic system. Using the cationic dye JC-1, early apoptotic changes were assessed following 5 h of HNE and H(2)O(2) insult. Though HNE exposure resulted in approximately 50% cells to undergo early apoptotic changes, no such changes were observed in H(2)O(2) treated cells during this period. Furthermore, apoptosis, as determined by quantifying the DNA fragmentation, was apparent at a much earlier time period by HNE as opposed to H(2)O(2). Taken together, the results demonstrate the apoptotic potential of the lipid peroxidation end-products and suggest that H(2)O(2)-induced apoptosis may be mediated by these end-products in the lens epithelium.

Aldehydes↗

PubMed2002

Characterization of flow and transport processes within the unsaturated zone of Yucca Mountain, Nevada, under current and future climates.

This paper presents a large-scale modeling study characterizing fluid flow and tracer transport in the unsaturated zone of Yucca Mountain, Nevada, a potential repository site for storing high-level radioactive waste. The study has been conducted using a three-dimensional numerical model, which incorporates a wide variety of field data and takes into account the coupled processes of flow and transport in the highly heterogeneous, unsaturated fractured porous rock. The modeling approach is based on a dual-continuum formulation of coupled multiphase fluid and tracer transport through fractured porous rock. Various scenarios of current and future climate conditions and their effects on the unsaturated zone are evaluated to aid in the assessment of the proposed repository's system performance using different conceptual models. These models are calibrated against field-measured data. Model-predicted flow and transport processes under current and future climates are discussed.

Climate↗

PubMed2002

Identification and localization of multiple classic cadherins in developing rat limbic system.

Classic cadherins are multifunctional adhesion proteins that play roles in tissue histogenesis, neural differentiation, neurite outgrowth and synapse formation. Several lines of evidence suggest that classic cadherins may establish regional or laminar recognition cues by virtue of their differential expression and tight, and principally homophilic, cell adhesion. As a first step toward investigating the role this family plays in generating limbic system connectivity, we used RT-PCR to amplify type I and type II classic cadherins present in rat hippocampus during the principal period of synaptogenesis. We identified nine different cadherins, one of which, cadherin-9, is novel in hippocampus. Using in situ hybridization, we compared the cellular and regional distribution of five of the cadherins (N, 6, 8, 9 and 10) during the first two postnatal weeks in hippocampus, subiculum, entorhinal cortex, cingulate cortex, anterior thalamus, hypothalamus and amygdala. We find that each cadherin is differentially distributed in distinct, but highly overlapping fields that largely correspond to known anatomical boundaries and are often coordinately expressed in interconnected regions. For example, cadherin-6 expression defines CA1 and its principal target, the subiculum; cadherin-10 is differentially expressed in CA1 and CA3 in a manner correlating with the organization of interconnecting Schaffer collateral axons; and cadherin-9 shows a striking concentration in CA3. Some cadherin mRNAs are highly restricted to particular anatomical fields over the entire time course, while others are more broadly expressed and become concentrated within particular domains coincident with the timing of afferent ingrowth. Our data indicate that classic cadherins are sufficiently diverse and differentially distributed to support a role in cell surface recognition and adhesion during the formation of limbic system connectivity.

Amygdala↗